title: Antioxidant efficacy and adhesion rescue by a recombinant mussel foot protein-6.
abstract: Mytilus foot protein type 6 (mfp-6) is crucial for maintaining the reducing conditions needed for optimal wet adhesion in marine mussels. In this report, we describe the expression and production of a recombinant Mytilus californianus foot protein type 6 variant 1 (rmfp-6.1) fused with a hexahistidine affinity tag in Escherichia coli and its purification by affinity chromatography. Recombinant mfp-6 showed high purification yields of 5-6 mg L(-1) cell culture and excellent solubility in low pH buffers that retard oxidation of its many thiol groups. Purified rmfp-6.1 protein showed high 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity when compared with vitamin C. Using the highly sensitive surface forces apparatus (SFA) technique to measure interfacial surface forces in the nano-Newton range, we show that rmfp-6.1 is also able to rescue the oxidation-dependent adhesion loss of mussel foot protein 3 (mfp-3) at pH 3. The adhesion rescue is related to a reduction of dopaquinone back to 3,4-dihydroxyphenyl-l-alanine in mfp-3, which is the reverse reaction observed during the detrimental enzymatic browning process in fruits and vegetables. Broadly viewed, rmfp-6.1 has potential as a versatile antioxidant for applications ranging from personal products to antispoilants for perishable foods during processing and storage. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:1587-1593, 2013.
authors.affiliation: Dept. of Molecular, Cellular and Developmental Biology, University of California, Santa Barbara, CA, 93106.
authors.forename: Sascha C T
authors.initials: SC
authors.lastname: Nicklisch
authors.forename: Saurabh
authors.initials: S
authors.lastname: Das
authors.forename: Nadine R
authors.initials: NR
authors.lastname: Martinez Rodriguez
authors.forename: J Herbert
authors.initials: JH
authors.lastname: Waite
authors.forename: Jacob N
authors.initials: JN
authors.lastname: Israelachvili
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000975:Antioxidants / Q000737:chemistry / Q000302:isolation & purification*; D009124:Muscle Proteins / Q000302:isolation & purification*; D049878:Mytilus / Q000737:chemistry* / Q000235:genetics; D010084:Oxidation-Reduction; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D000975:Antioxidants; D009124:Muscle Proteins; D011994:Recombinant Proteins
keywords: DOPA oxidation; adhesion rescue; antioxidant; recombinant mussel protein; thiol